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Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Published on: July 28, 2017

DNA labeling by isolated wheat chloroplasts.

¹The Biotechnology Centre, University of Cambridge, Cambridge, UK.

Methods in Molecular Biology (Clifton, N.J.)

|March 23, 2011

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Wheat leaf cells replicate chloroplast DNA in a specific 1-4 cm region from the base in young seedlings. Isolated chloroplasts from this zone can synthesize DNA in vitro, indicating active DNA replication.

Area of Science:

Plant Biology
Molecular Biology
Genetics

Background:

Wheat plant growth originates from a basal meristem.
Young wheat leaves exhibit a developmental gradient, with cells aging from tip to base.
Chloroplast DNA replication is a key process for chloroplast biogenesis and function.

Purpose of the Study:

To investigate the specific location and timing of chloroplast DNA replication in developing wheat leaves.
To confirm the capacity for in vitro DNA synthesis in isolated chloroplasts from a defined leaf region.

Main Methods:

Analyzing cellular development and DNA replication across different regions of young wheat leaves.
Isolating chloroplasts from a specific 1-4 cm segment of wheat seedlings (up to 10 days old).

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Last Updated: Jun 3, 2026

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Published on: July 28, 2017

Analysis of Protein Import into Chloroplasts Isolated from Stressed Plants

Analysis of Protein Import into Chloroplasts Isolated from Stressed Plants

Published on: November 1, 2016

Incubating isolated chloroplasts with radioactively labeled DNA precursors to assess in vitro DNA synthesis.

Main Results:

Chloroplast DNA replication was localized to a specific region (1-4 cm from the leaf base) in wheat seedlings up to 10 days old.
Isolated chloroplasts from this identified leaf region demonstrated the ability to produce radioactively labeled DNA in vitro.

Conclusions:

The 1-4 cm region from the leaf base in young wheat seedlings is a primary site of active chloroplast DNA replication.
Isolated wheat chloroplasts possess the enzymatic machinery for DNA synthesis, supporting in vitro replication studies.